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Generation of a vector suite for protein solubility screening
Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization o...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934309/ https://www.ncbi.nlm.nih.gov/pubmed/24616717 http://dx.doi.org/10.3389/fmicb.2014.00067 |
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author | Correa, Agustín Ortega, Claudia Obal, Gonzalo Alzari, Pedro Vincentelli, Renaud Oppezzo, Pablo |
author_facet | Correa, Agustín Ortega, Claudia Obal, Gonzalo Alzari, Pedro Vincentelli, Renaud Oppezzo, Pablo |
author_sort | Correa, Agustín |
collection | PubMed |
description | Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning and screening methods. Here we describe a new versatile vector suite dedicated to the expression improvement of recombinant proteins (RP) with solubility problems. This vector suite allows the parallel cloning of the same PCR product into the 12 different expression vectors evaluating protein expression under different promoter strength, different fusion tags as well as different solubility enhancer proteins. Additionally, we propose the use of a new fusion protein which appears to be a useful solubility enhancer. Above all we propose in this work an economic and useful vector suite to fast track the solubility of different RP. We also propose a new solubility enhancer protein that can be included in the evaluation of the expression of RP that are insoluble in classical expression conditions. |
format | Online Article Text |
id | pubmed-3934309 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-39343092014-03-10 Generation of a vector suite for protein solubility screening Correa, Agustín Ortega, Claudia Obal, Gonzalo Alzari, Pedro Vincentelli, Renaud Oppezzo, Pablo Front Microbiol Microbiology Recombinant protein expression has become an invaluable tool for academic and biotechnological projects. With the use of high-throughput screening technologies for soluble protein production, uncountable target proteins have been produced in a soluble and homogeneous state enabling the realization of further studies. Evaluation of hundreds conditions requires the use of high-throughput cloning and screening methods. Here we describe a new versatile vector suite dedicated to the expression improvement of recombinant proteins (RP) with solubility problems. This vector suite allows the parallel cloning of the same PCR product into the 12 different expression vectors evaluating protein expression under different promoter strength, different fusion tags as well as different solubility enhancer proteins. Additionally, we propose the use of a new fusion protein which appears to be a useful solubility enhancer. Above all we propose in this work an economic and useful vector suite to fast track the solubility of different RP. We also propose a new solubility enhancer protein that can be included in the evaluation of the expression of RP that are insoluble in classical expression conditions. Frontiers Media S.A. 2014-02-25 /pmc/articles/PMC3934309/ /pubmed/24616717 http://dx.doi.org/10.3389/fmicb.2014.00067 Text en Copyright © 2014 Correa, Ortega, Obal, Alzari, Vincentelli and Oppezzo. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Correa, Agustín Ortega, Claudia Obal, Gonzalo Alzari, Pedro Vincentelli, Renaud Oppezzo, Pablo Generation of a vector suite for protein solubility screening |
title | Generation of a vector suite for protein solubility screening |
title_full | Generation of a vector suite for protein solubility screening |
title_fullStr | Generation of a vector suite for protein solubility screening |
title_full_unstemmed | Generation of a vector suite for protein solubility screening |
title_short | Generation of a vector suite for protein solubility screening |
title_sort | generation of a vector suite for protein solubility screening |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934309/ https://www.ncbi.nlm.nih.gov/pubmed/24616717 http://dx.doi.org/10.3389/fmicb.2014.00067 |
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